CALLUS INDUCTION OF CACAO CLONE SULAWESI 1 ON VARIOUS CONCENTRATIONS OF  2,4 -D AND COCONUT WATER VIA IN VITRO CULTURE

Cacao is one of important plantation crops grouped in the Sterculiaceae family. Sulawesi is the main area of cacao production and has a number of superior clones, such as Sulawesi 1 and Sulawesi 2. Based on data in 2012/2014 cacao production to consumption reached 174,000 tons, while in 2013/2014 was projected a deficit of 115,000 tonnes. Nonetheless, cacao agribusiness in Indonesia is still facing complex problems, among others gardener productivity is still low due to borer attacks cacao, the quality of products and the number is still low and still not optimal development of cacao products and providing superior amount of cacao seedlings. The primary problem of cacao production recently is low productivity. The main cause of low cacao productivity in Central Sulawesi is the use of inferior clones. To enhance cacao productivity, it is crucial to use cacao clones having high genetic potential via tissue culture or micropropagation techniques. The aim of this experiment was to assess the effect of different concentrations of 2,4-D and coconut water on the growth of cacao callus via in vitro culture. This experiment used Completely Randomozed Block Design in factorial patteren with treatments tested namely 2,4-D and coconut water concentrations. The concentrations of 2,4-D tested including 1 ppm, 2 ppm and 3 ppm, whilst coconut water concentrations tested consisting of 10%, 15% and 20%, and therefore there were 3 x 3 = 9 treatment combinations. Each treatment utilized 4 replications; and each unit combination used 5 explants (staminodia). Results of this experiment indicated that the addition of 3 ppm 2,4-D and 10% coconut water had a highly significant effect on the callus color 4 weeks after culture. The addition of 3 ppm 2,4-D in culture media showed a significant effect on callus color 4 weeks after culture, but had an insignificant effect on the callus formation, callus color 8 weeks after culture an callus texture. Supplementation of 20% coconut water had a significant effect on callus texture 8 weeks after culture, whilst the addition of 10% coconut water showed a significant effect on callus color 4 weeks after culture.

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PENGARUH AIR KELAPA TERHADAP MULTIPLIKASI TUNAS IN VITRO, PRODUKSI RIMPANG, DAN KANDUNGAN XANTHORRHIZOL TEMULAWAK DI LAPANGAN

Jurnal Penelitian Tanaman Industri ◽

10.21082/jlittri.v18n3.2012.125-134 ◽

2020 ◽

Vol 18 (3) ◽

pp. 125 ◽

Cited By ~ 2

Author(s):

NATALINI NOVA KRISTINA ◽

SITTI FATIMAH SYAHID

Keyword(s):

In Vitro Culture ◽

Growth Medium ◽

Growth Regulator ◽

Block Design ◽

Hplc Method ◽

Coconut Water ◽

Growth And Yield ◽

Chemical Content ◽

Curcuma Xanthorrhiza

ABSTRAK Langkah antisipatif pemenuhan kebutuhan massal benih temulawak dilakukan dengan perbanyakan secara in vitro menggunakan medium tumbuh yang murah mengandung air kelapa. Penelitian bertujuan untuk menganalisis kandungan kimia air kelapa dan peranannya dalam multi- plikasi tunas temulawak in vitro, serta pengaruhnya terhadap produksi rimpang dan kandungan xanthorrizol. Penelitian dilakukan mulai Mei 2009 sampai Agustus 2010 di Laboratorium dan Kebun Percobaan Balai Penelitian Tanaman Rempah dan Obat; serta Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian. Air kelapa yang digunakan berasal dari kelapa muda (7-8 bulan) dan kelapa tua berumur (10-12 bulan). Penelitian dilakukan secara bertahap, terdiri atas 4 kegiatan. Pertama, analisis zat pengatur tumbuh, vitamin dan mineral dalam air kelapa menggunakan metode HPLC. Kedua, pengaruh konsentrasi air kelapa (0, 5, 10, 15, 20, dan 25%) terhadap multiplikasi tunas temulawak in vitro. Kegiatan dirancang secara acak kelompok, 3 ulangan. Pengamatan meliputi parameter pertumbuhan. Ketiga, aklimatisasi dan kandungan klorofil tanaman hasil in vitro. Keempat, pertumbuhan dan produksi rimpang benih temulawak in vitro dalam pot berisi media tanah + pasir dan analisis kandungan xanthorrizolnya. Rancangan penelitian acak kelompok, 3 ulangan, dan parameter pengamatan karakter pertumbuhan, produksi rimpang, dan kandungan xanthorrizol. Hasil penelitian menunjukkan bahwa air kelapa mengandung kinetin, zeatin, auksin, vitamin, mineral dan sumber karbon yang berguna untuk multiplikasi tunas in vitro. Kandungan kimia air kelapa muda lebih tinggi dibanding air kelapa tua. Medium tumbuh mengandung air kelapa 15% terbaik dalam merangsang pertum- buhan tunas in vitro (rata-rata 4,6 jumlah tunas per botol selama periode awal pertumbuhan (8 minggu) sehingga dijadikan sebagai standar perba- nyakan. Bibit temulawak hasil perbanyakan in vitro tumbuh baik (72%) pada masa aklimatisasi, walaupun sebagian kecil ada yang menguning. Kandungan klorofil a, b, dan total klorofil temulawak asal kultur in vitro lebih tinggi dibandingkan dengan yang konvensional, dan bentuk rimpangnya normal. Poduksi rimpang generasi awal (Vo) mencapai rata- rata 320,2g, lebih rendah dibandingkan dengan rimpang konvensional (800,5g). Kandungan xanthorrhizol temulawak hasil kultur in vitro lebih rendah dibandingkan rimpang konvensional. Hasil penelitian mengindi- kasikan potensi air kelapa sebagai zat pengatur tumbuh alami pada temulawak in vitro. Kata kunci: air kelapa, Curcuma xanthorrhiza, in vitro, xanthorrhizol, hasilABSTRACT Anticipated step for Java turmeric seed massal fulfillment was conducted by in vitro using cheap growth medium enriched with coconut water. The aim of the research was to analyse the chemical content of coconut water and its role on java turmeric micropropagation in vitro and their effect on yield and xanthorrhizol content. The experiement was conducted from May 2009 to August 2009 at Indonesian Spices and Medicinal Research Institute and Indonsian Center for Agricultural Post Harvest Research and Development. The coconut water used comes from young coconut (7-8 months) and old coconut (10-12 months). The research consisted of four steps. First, analysis of growth regulator, vitamin and sucrose from coconut water using HPLC method. Second, the effect of several concentration od water coconut: 0, 5, 10, 15, 20, and 25% on in vitro multiplication. The experiment was arranged in completely block design with three replicates. The parameters observed were growth of culture during in vitro. Third, acclimatization and chlorophyll content of plant derived from in vitro and fourth, growth, and yield of java turmeric seed on pot containing soil + sand as growth medium and xanthorrhizol analysis. The experiment was arranged in completely block design with three replicates. The parameters observed were growth characters, yield and xnthorrhizol content. Result showed that coconut water contain kinetin, zeatin, auksin, vitamin, mineral and carbon source which used for in vitro shoots multiplication. The chemical of young coconut water was higher than old coconut. The growth medium enriched with 15 % coconut water gave the best result on inducing shoots in vitro (average 4.6 shoots/bottle during 8 weeks culture), so it’s used as multiplication standard. Java turmeric seed from in vitro culture grew well (72%) on acclimatization. Although, some of them were greenish.The content of a, b, and total chlorophyll of java ginger from in vitro culture was higher than conventional rhizome and have a normal rhizome. The production on Vo (plantlet generation) around 320.2 g/plant, is lower than conventional rhyzome (800.5 g). Xanthorhizol and essential oil content of Java turmeric from in vitro seed were lower than conventional rhyzome. Result research indicated potency of the coconut water as a nature growth regulator in vitro. Key words: coconut water, Curcuma xanthorrhiza, in vitro, growth, xanthorrhizol, yield

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75 Improvement of Developmental Competence of Bovine In Vitro-Produced Embryos by Using Charcoal:Dextran-Stripped Fetal Bovine Serum on In Vitro Culture Media

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab75 ◽

2018 ◽

Vol 30 (1) ◽

pp. 176

Author(s):

A. Mesalam ◽

R. Kong ◽

B.-H. Choi ◽

K.-L. Lee ◽

B.-Y. Park ◽

...

Keyword(s):

In Vitro Culture ◽

Bovine Serum ◽

Culture Media ◽

Fetal Bovine Serum ◽

Developmental Competence ◽

Mitochondrial Activity ◽

Mrna Levels ◽

Fetal Bovine

Serum has widely been used as a main supplement to embryo in vitro culture media as it contains embryotrophic factors. Charcoal:dextran treatment of fetal bovine serum (FBS) removes lipophilic chemicals and certain steroid hormones and growth factors. The objective of this study was to investigate the effects of charcoal:dextran-stripped fetal bovine serum (CDS FBS) and heat-inactivated FBS (HI FBS) in embryo culture medium (SOF-BE1 medium supplemented with 10% of serum) on their ability to support in vitro development of bovine embryos. The developmental ability and quality of bovine embryos were determined by assessing their cell number, lipid content, mitochondrial activity, gene expression, and cryo-tolerance. The experiment was conducted in 6 replicates (350 oocytes per group). The differences in embryo development, integrated optical intensity, and expression levels of the various genes between experimental groups were analysed by one-way ANOVA. Duncan’s multiple range tests were used to test the differences between the treatments. The level of statistical significance was set at P < 0.05. The percentages of embryos that underwent cleavage and formed a blastocyst were significantly (P < 0.05) higher in medium containing CDS FBS than in medium containing HI FBS (42.84 ± 0.78% v. 36.85 ± 0.89%, respectively). The total number of cells per Day 8 blastocyst was not significantly different (P > 0.05) between the CDS FBS group (208.40 ± 14.77) and the HI FBS group (195.11 ± 19.15). Furthermore, the beneficial effects of CDS FBS on embryos were associated with a significantly increased mitochondrial activity, as identified by MitoTracker Green, and reduced intracellular lipid content, as identified by Nile red staining, which increased their cryo-tolerance. The post-thaw survival rate of blastocysts was significantly (P < 0.05) higher after 24 h in the CDS FBS than in the HI FBS group (85.33 ± 4.84% v. 68.67 ± 1.20%). Quantitative reverse transcription PCR showed that the mRNA levels of lipid metabolism-related genes, acyl-CoA synthetase long-chain family member 3, acyl-coenzyme A dehydrogenase long-chain, and the cholesterol metabolism related gene hydroxymethylglutaryl-CoA reductase were significantly increased upon culture with CDS FBS. Moreover, the mRNA levels of survival gene sirtuin 1, antioxidant gene superoxide dismutase 2, and anti-apoptotic associated gene B-cell lymphoma 2 in frozen–thawed blastocysts were significantly (P < 0.05) higher in the CDS FBS group than in the HI FBS group; however, the mRNA level of the pro-apoptotic gene BCL2-associated X protein was significantly reduced. In conclusion, these data suggest that supplementation of in vitro culture medium with CDS FBS improves in vitro bovine embryo developmental competence and the quality of blastocysts in terms of their crytolerance and gene expression. This research was supported by grant from the Next-Generation BiogGeen21 (No. PJ01107703), IPET (No. 315017-5 and 117029-3), Allergy free cat (Co.. Felix Pets), BK21plus, and KGSP.

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Lippia graveolens (Lamiales: Verbenaceae) and Oryganum vulgare (Lamiales: Lamiaceae) obtained by means of the in vitro propagation technique

RECUBRIMIENTO COMESTIBLE CON BASE DE PIÑON MEXICANO (Jatropha curcas) SOBRE LA CALIDAD DE CHAYOTE MINIMAMENTE PROCESADO ◽

10.32854/agrop.v14i11.2177 ◽

2021 ◽

Author(s):

María A. Aguilar Morales ◽

Armandina De la Cruz Olvera ◽

E. Archundia-Garduño ◽

Rosy G. Cruz Monterrosa ◽

Mayra Díaz-Ramírez ◽

...

Keyword(s):

Activated Carbon ◽

Culture Media ◽

Callus Formation ◽

Block Design ◽

Leaf Explants ◽

Basal Medium ◽

Culture Technique ◽

Ms Medium ◽

Antioxidant Agents

Objective: The objective of this study was to establish the method of propagation of Oryganum vulgare and Lippia graveolens employing a plant tissue culture technique that decreased the phenolization percentages and increased the multiplication coefficients. Design/ methodology/ approach: The in vitro germination percentage was evaluated in both MS and MS medium + activated carbon. Microcuttings (small shoots) of both species were established in base medium added with different antioxidant agents to decrease the phenolization of explants; the treatments were arranged in a completely randomized block design. For the propagation phase, a completely randomized factorial design was used, where the auxin/cytokinin phytoregulators, type of explants (axillary buds and leaves), and the species (Lippia graveolens and Oryganum vulgare) were considered as factors. Results: maximum germination (63.3% ±12.5) was obtained on day 15 in both culture media for L. graveolens and O. vulgare. The use of antioxidant agents mainly activated carbon, increased the in vitro establishment and activation of vegetative buds in both species by up to 90%. There were significant differences in the variables evaluated regarding the treatments, the explant, and the species in the multiplication phase. The combination 1.0/ 0.5 mg L-1 BA/AIB induces callus formation for both species. When used as leaf explants, callus formation was potentiated. Study Limitations / Implications: The results presented are advances from a long-term experiment. Findings/conclusions: The germination of L. graveolens seeds can be achieved in MS medium after 15 days. Microcuttings of both L. graveolens and O. vulgare were successfully established in MS basal medium enriched with 1 g L-1 charcoal that showed low oxidation percentages and induced up to 90% the production of shoots in the explants. The mixture of 1.0/0.5 mg L-1 BA/AIB induces callus formation for both species; when this medium is in contact with leaves as an explant, its formation is potentiated, achieving diameters up to 15 mm. In order to achieve the induction of shoots and roots, buds should be established in MS medium enriched with 0.5 mg L-1 IBA for both species; this mixture encreased the multiplication coefficients

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Studi Kualitas Regeneran Phalaenopsis Hasil Kultur In Vitro dari Eksplan Tangkai Infloresen, Tunas Pucuk, dan Empulur (The Quality Study of Phalaenopsis Regenerants from In Vitro Propagation of Inflorescence, Shoot Tip, and Pith Explants)

Jurnal Hortikultura ◽

10.21082/jhort.v28n1.2018.p13-24 ◽

2019 ◽

Vol 28 (1) ◽

pp. 13

Author(s):

Dewi Pramanik ◽

Herni Shintiavira ◽

Budi Winarto

Keyword(s):

Clonal Propagation ◽

Callus Formation ◽

Stomatal Density ◽

Average Length ◽

Block Design ◽

Shoot Tips ◽

The Impact ◽

In Vitro Clonal Propagation

Anggrek Phalaenopsis memiliki nilai komersial yang tinggi, karena keindahannya dapat dinikmati sepanjang tahun. Hal tersebut berdampak pada kebutuhan benih tanaman yang semakin meningkat. Salah satu cara penyediaan benih secara massal adalah melalui perbanyakan klonal secara in vitro sehingga perlu dilakukan studi kualitas regeneran hasil perbanyakan klonal untuk menjamin ketersediaan benih dengan kualitas baik. Penelitian bertujuan menguji kualitas regeneran yang dihasilkan dari perbanyakan klonal secara in vitro beberapa varietas Phalaenopsis dengan menggunakan eksplan yang berbeda. Penelitian dilaksanakan di Laboratorium Kultur Jaringan, Kebun Percobaan Segunung, Balai Penelitian Tanaman Hias (Balithi) sejak bulan Januari 2014 hingga Mei 2015. Penelitian menggunakan dua faktor, yaitu varietas (Ayu Lestari, Ayu Pratiwi, dan Karindra) dan jenis eksplan (tangkai infloresen, tunas pucuk, dan empulur). Percobaan disusun menggunakan rancangan acak kelompok pola faktorial dan setiap perlakuan diulang tiga kali. Hasil penelitian menunjukan tidak terjadi interaksi yang nyata antara faktor jenis eksplan dan varietas yang diujikan pada semua tahap percobaan. Respon terbaik diperoleh pada eksplan empulur dengan 42,85% eksplan berhasil membentuk kalus pada minggu ke-8 dan hampir 100% kalus tersebut dapat beregenerasi menjadi tunas pada minggu ke-24 dengan tingkat multiplikasi tunas 1,87 kali. Pada minggu ke-32 terbentuk rata-rata 3,13 daun per planlet dengan 2,47 cm panjang daun, 1,36 cm lebar daun, 1,52 akar per planlet, dan panjang akar per planlet mencapai 1,26 cm. Kerapatan stomata memiliki korelasi negatif dengan tingkat abnormalitas planlet. Planlet dengan kerapatan stomata tertinggi dan abnormalitas yang rendah diperoleh pada var. Karindra dan planlet yang berasal dari eksplan empulur dan tunas pucuk. Setelah 8 minggu tahap aklimatisasi, tingkat keberhasilan hidup tertinggi (92%) diperoleh pada tunas yang berasal dari eksplan empulur. Penelitian membuktikan bahwa perbedaan varietas tidak memiliki pengaruh nyata pada tingkat abnormalitas regeneran dan dari eksplan empulur diperoleh jumlah regeneran tertinggi dengan kualitas baik (tingkat abnormalitas rendah).KeywordsKultur jaringan; Kualitas regeneran; Phalaenopsis; Jenis eksplantAbstractPhalaenopsis orchids have a high commercial value, because of its beauty and it can be enjoyed throughout the year. This condition gives the impact on the increasing demand of the seeds. One of the ways of providing mass seeds is through in vitro clonal propagation. However, it is necessary to study the quality of regenerants of clonal propagation products to ensure the availability of qualified seeds. The aimed of this study was to test the quality of regenerants obtained from in vitro clonal propagation of Phalaenopsis using inflorescence stalk, shoot tips, and pith explants. This research was conducted at Tissue Culture Laboratory, Segunung Experimental Station, Indonesian Ornamental Crops Research Institute (IOCRI) from January 2014 to May 2015. The study used two treatments, varieties (Ayu Lestari, Ayu Pratiwi, and Karindra) and type of explant (inflorescence stalk, shoot tips, and pith). Experiments were prepared using a randomized complete block design with two factors and each treatment was replicated three times. The results showed there were no significant interaction between types of explants and varieties tested in all experiment stages. The best response was obtained using pith explants with 42.85% callus formation in the week eighth and nearly 100% callus can regenerate into shoots at week 24th with the rate of shoot multiplication up to 1.87 times. At week 32th the cultures formed planlets with an average number of leaves of 3.13 and an average size of 2.47 cm x 1.36 cm (length x width) and an average number of roots of 1.52 with average length reached 1.26 cm. Stomatal density has negative correlation with plantlet abnormality rate. Plantlets with the highest stomatal density and low abnormality were obtained in var. Karindra and plantlet derived from explant pith and shoot buds. After 8 weeks of acclimatization stage, the highest survival rate (92%) was obtained on the shoot originating from pith explant. This study proved that varietal differences did not have a significant effect on regenerant abnormalities, and the highest number of regenerant with good quality (low abnormality rate) was obtained from pith explant.

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Kualitas Semen Cair Kambing Boer Berbahan Pengencer Air Kelapa Muda Varietas Viridis Setelah Simpan Dingin

Jurnal Ilmu dan Teknologi Peternakan Tropis ◽

10.33772/jitro.v6i1.5449 ◽

2019 ◽

Vol 6 (1) ◽

pp. 78

Author(s):

Muhammad Ade Salim ◽

Muhammad Nur Ihsan ◽

Nur Isnaini ◽

Trinil Susilawati

Keyword(s):

Semen Quality ◽

Block Design ◽

Animal Husbandry ◽

Egg Yolk ◽

Coconut Water ◽

Boer Goat ◽

Randomized Block Design ◽

Frozen Semen ◽

Artificial Vagina

ABSTRAKAir kelapa muda varietas viridisdapat dijadikan pengencer aletrnatif semen cair bagi program IB di daerah minim sarana semen beku. Tujuan penelitian ini untuk menguji pengaruh penggunaan air kelapa muda viridissebagai bahan pengencer terhadap kualitas semen cair kambing Boer setelah didinginkan. Dilaksanakanselama 3 bulan di Laboratorium Fakultas Peternakan UBUnit SumberSekar,Malang. Metodenya yaitu eksperimen. Semen dari 3 pejantan Boer umur 3-5 tahun, dikoleksi seminggu sekali dengan VB. Air kelapa mudaviridis umur 5-7 bulan serta tris aminomethane sebagai kontrol. Didesain menggunakan Rancangan Acak Kelompok (RAK) dengan 2 perlakuan yaitu P0 (tris aminomethane + 10% KT) dan P1 (air kelapa muda viridis + 10% KT) masing-masing diulang 10 kali. Data dianalisis dengan analisis Ragam (Anova) dengan software Genstat 18. Variabelnya yaitu motilitas individu, viabilitas dan abnormalitas. Hasil penelitian yaitu motilitas individu pada P1bertahan sampai 4 hari (40,5± 24,3%), viabilitas terbaik sampai hari ke-5 (42±24,6%), abnormalitas terendah di hari ke-7(1,31± 0,6). Kesimpulannya, Pengencer air kelapa muda viridis dapat mempertahankan kualitas semen cair kambing Boer selama 4 hari untuk motilitas dan 5 hari untuk viabilitas.Kata Kunci:pengencer, air kelapa, varietas viridisABSTRACTYoung viridis coconut water could be used as an alternative to liquid semen diluent for artificial insemination program in the area with limited facility for frozen semen production. This study evaluated the use of young coconut water as a diluent on liquid semen quality of Boer goat after cold storage. This study was carried out for 3 months at Sumber Sekar Laboratory, Faculty of Animal Husbandry, University of Brawijaya, Malang. The semen was collected from 3 Boer bucks aged at 3 to 5 years old. The semen collection was done once a week with the aid of artificial vagina. The diluents used were young Viridis coconut (5 to 7 months old) and tris aminomethane. The method used was an experiment in a randomized block design with 2 treatments and 10 replicates. The treatments used were T0: tris aminomethane + 10% egg yolk (control) and T1: young Viridis coconut water + 10% egg yolk. Data were analyzed by analysis of variance using Genstat 18 software. The variables measured were sperm individual motility, viability, and abnormality. The results showed that the sperm individual motility in T1 survived up to 4 days (40.5± 24.3%), the best viability at 5 days (42.0±24.6%), while the lowest abnormality at 7 days (1.31±0.6). It could be concluded that: 1. Tris aminomethane diluent has higher quality with the storage length up to 9 days, 2. Young Viridis coconut water diluent could preserve liquid semen quality of Boer goat up to 4 days for sperm motility and 5 days for sperm viability.Keywords: diluents, coconut water, viridis variety

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Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽

10.30598/a.v1i1.293 ◽

2018 ◽

Vol 1 (1) ◽

Author(s):

S. Tuhuteru ◽

Meity L Hehanussa ◽

Simon H.T Raharjo

Keyword(s):

Culture Medium ◽

Culture Media ◽

Water Concentration ◽

Medium Composition ◽

Coconut Water ◽

Orchid Species ◽

Randomized Design ◽

Wet Weight ◽

Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media. The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids. Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

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Rheological and Microstructural Features of Plant Culture Media Doped with Biopolymers: Influence on the Growth and Physiological Responses of In Vitro-Grown Shoots of Thymus lotocephalus

Polysaccharides ◽

10.3390/polysaccharides2020032 ◽

2021 ◽

Vol 2 (2) ◽

pp. 538-553

Author(s):

Natacha Coelho ◽

Alexandra Filipe ◽

Bruno Medronho ◽

Solange Magalhães ◽

Carla Vitorino ◽

...

Keyword(s):

In Vitro Culture ◽

Culture Medium ◽

Culture Media ◽

Average Pore Size ◽

Physiological Responses ◽

Gum Arabic ◽

Shoot Elongation ◽

Hydroxyethyl Cellulose ◽

Plant Culture

In vitro culture is an important biotechnological tool in plant research and an appropriate culture media is a key for a successful plant development under in vitro conditions. The use of natural compounds to improve culture media has been growing and biopolymers are interesting alternatives to synthetic compounds due to their low toxicity, biodegradability, renewability, and availability. In the present study, different culture media containing one biopolymer (chitosan, gum arabic) or a biopolymer derivative [hydroxyethyl cellulose (HEC), carboxymethyl cellulose (CMC)], at 100 or 1000 mg L−1, were tested regarding their influence on the growth and physiological responses of Thymus lotocephalus in vitro culture. Cellulose-based biopolymers (HEC and CMC) and gum arabic were used for the first time in plant culture media. The results showed that CMC at 100 mg L−1 significantly improved shoot elongation while chitosan, at the highest concentration, was detrimental to T. lotocephalus. Concerning only the evaluated physiological parameters, all tested biopolymers and biopolymer derivatives are safe to plants as there was no evidence of stress-induced changes on T. lotocephalus. The rheological and microstructural features of the culture media were assessed to understand how the biopolymers and biopolymer derivatives added to the culture medium could influence shoot growth. As expected, all media presented a gel-like behaviour with minor differences in the complex viscosity at the beginning of the culture period. Most media showed increased viscosity overtime. The surface area increased with the addition of biopolymers and biopolymer derivatives to the culture media and the average pore size was considerably lower for CMC at 100 mg L−1. The smaller pores of this medium might be related to a more efficient nutrients and water uptake by T. lotocephalus shoots, leading to a significant improvement in shoot elongation. In short, this study demonstrated that the different types of biopolymers and biopolymer derivatives added to culture medium can modify their microstructure and at the right concentrations, are harmless to T. lotocephalus shoots growing in vitro, and that CMC improves shoot length.

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Somaclonal variation on in vitro callus culture potato cultivars

Horticultura Brasileira ◽

10.1590/s0102-05362004000200027 ◽

2004 ◽

Vol 22 (2) ◽

pp. 300-304 ◽

Cited By ~ 28

Author(s):

Patricia N. Bordallo ◽

Derly H. Silva ◽

José Maria ◽

Cosme D. Cruz ◽

Elizabeth P. Fontes

Keyword(s):

Somaclonal Variation ◽

Culture Media ◽

Callus Formation ◽

Potato Cultivars ◽

Synthetic Seed ◽

Arbitrary Sequence ◽

Stem Explants ◽

Factors Affecting ◽

High Level

Synthetic seeds can be an alternative for those species in which botanical seeds are not viable. One of the major problems of in vitro plant cultivation is the high level of somaclonal variation. The most common factors affecting somaclonal variation are genotype, explant source, in vitro period and cultivation conditions in which the culture is established. In this work, calli were induced using leaf and stem explants of the commercial potato cultivars Achat, Baraka, Baronesa, Bintje, and Contenda in MS culture media supplemented with 1.65 mM of picloram and 11.5 mM of 2,4-D. Seventy and 90 days after induction, DNA samples of 40 calli were compared concerning the effects of the two explant (leaf and stem) and two growth regulator sources on five potatoes cultivars. A total of 20 arbitrary sequence primers were evaluated. The RAPD pattern generated by these primers suggested a high percentage of polymorphic fragments among the five genotypes, indicating a high level of genetic variation among cultivars. Cultivar Baronesa showed the highest number of polymorphic fragments for all treatments. The cultivar Contenda showed the smallest somaclonal variation, for most of the treatments, except for the treatment which consisted of stem explants, picloram (1.65 mM) application, and a 70-day period of callus formation. 'Contenda' is, therefore, the most suitable cultivar for synthetic seed production.

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Biochemical markers for pregnancy in the spent culture medium of in vitro produced bovine embryos

Biology of Reproduction ◽

10.1093/biolre/ioab095 ◽

2021 ◽

Author(s):

Gabriela de Oliveira Fernandes ◽

Marcella Pecora Milazzotto ◽

Andrei Antonioni Guedes Fidelis ◽

Taynan Stonoga Kawamoto ◽

Ligiane de Oliveira Leme ◽

...

Keyword(s):

Mass Spectrometry ◽

Biochemical Markers ◽

Culture Medium ◽

Culture Media ◽

Ionization Mass ◽

Metabolic Profiles ◽

Bovine Embryos ◽

The Media

Abstract The present study aimed to identify biomarkers to assess the quality of in vitro produced (IVP) bovine embryos in the culture media. IVP embryos on Day (D) 5 of development were transferred to individual drops, where they were maintained for the last 48 h of culture. Thereafter, the medium was collected and the embryos were transferred to the recipients. After pregnancy diagnosis, the media were grouped into the pregnant and nonpregnant groups. The metabolic profiles of the media were analyzed via electrospray ionization mass spectrometry, and the concentrations of pyruvate, lactate, and glutamate were assessed using fluorimetry. The spectrometric profile revealed that the media from embryos from the pregnant group presented a higher signal intensity compared to that of the nonpregnant group; the ions 156.13 Da [M + H]+, 444.33 Da [M + H]+, and 305.97 Da [M + H]+ were identified as biomarkers. Spent culture medium from expanded blastocysts (Bx) that established pregnancy had a greater concentration of pyruvate (p = 0.0174) and lesser concentration of lactate (p = 0.042) than spent culture medium from Bx that did not establish pregnancy. Moreover, pyruvate in the culture media of Bx can predict pregnancy with 90.9% sensitivity and 75% specificity. In conclusion, we identified markers in the culture media that helped in assessing the most viable IVP embryos with a greater potential to establish pregnancy.

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In vitro culture as an aid to conservation of indigenous ferns: Diplazium proliferum

International Journal of Plant Biology ◽

10.4081/pb.2015.6020 ◽

2015 ◽

Vol 6 (1) ◽

Author(s):

Zubeir M. Golamaully ◽

Vishwakalyan Bhoyroo ◽

Nadeem Nazurally ◽

Vineshwar Gopal

Keyword(s):

Growth Rate ◽

In Vitro Culture ◽

Mercuric Chloride ◽

Rare Species ◽

Vascular Plants ◽

Culture Media ◽

Fungal Contamination ◽

Increase Growth Rate ◽

Growing Population

With the ever growing population and economic needs of Mauritius, the flora of Mauritius has never been in more danger and one group of vascular plants is even more in peril; ferns. Diplazium proliferum is indigenous to the Mascarene region and is considered as a rare species in Mauritius. The need to develop a tested in vitro propagation protocol is a must to protect the biodiversity of Mauritius. This experiment was geared towards the establishment of a proper sterilization technique and the effect of 6-benzylaminopurine (BAP) and light on in vitro culture of this fern. Sterilization with 0.05% Mercuric chloride was effective to eliminate fungal contamination and allow germination of spores. Culture media supplemented with BAP did not significantly increase growth rate of both gametophytes and sporophytes of D. proliferum. Present results suggest efficient sterilization methods to be a crucial stage for successful in vitro regeneration of ferns. The established protocol will be used as an optimized baseline protocol for the propagation of other indigenous ferns.

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